P250 high capacity slide scanner

Manufactured by PerkinElmer

The P250 High Capacity Slide Scanner is a laboratory equipment designed for high-throughput digitization of microscope slides. It features a large capacity slide loader, enabling the automated scanning of multiple slides in a single session. The device utilizes advanced optics and imaging technology to capture high-resolution digital images of the slide contents.

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Lab products found in correlation

Neutral buffered formalin, by Thermo Fisher Scientific (2 mentions) Apoptag peroxidase in situ apoptosis detection kit, by Merck Group (2 mentions) Anti brdu antibody, by Cell Signaling Technology (1 mentions) Sp 2001, by Vector Laboratories (1 mentions) Phospho ser thr akt substrate, by Cell Signaling Technology (1 mentions) β catenin, by Cell Signaling Technology (1 mentions) Histo clear reagent, by National Diagnostics (1 mentions) Cleaved caspase 3 antibody, by Cell Signaling Technology (1 mentions) Buffered formalin, by Thermo Fisher Scientific (1 mentions)

8 protocols using p250 high capacity slide scanner

Quantitative Histopathological Analysis of Lungs

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Lungs were removed and fixed in 10% neutral buffered formalin (Fisher Scientific, Fair Lawn, NJ). Tissues were embedded with paraffin, sectioned at 5 μm thickness, and stained with haematoxylin and eosin stain. Five different lung sections per mouse were analyzed. Slides were scanned with a Perkin Elmer P250 High Capacity Slide Scanner (Waltham, Massachusetts) at 2,000 dots per inch (dpi). Digitized images were then analyzed using ImageJ software to calculate the total disease area occupied by granuloma and the percentage of lung surface affected by pneumonia as well as the number of infiltrates per lung. The total disease area for the entire lung section was calculated by adding the values for each lesion. The total percentage of diseased tissue was calculated by dividing the total disease area by the entire lung section and multiplying by 100, using image J software.

Prados-Rosales R., Carreño L., Cheng T., Blanc C., Weinrick B., Malek A., Lowary T.L., Baena A., Joe M., Bai Y., Kalscheuer R., Batista-Gonzalez A., Saavedra N.A., Sampedro L., Tomás J., Anguita J., Hung S.C., Tripathi A., Xu J., Glatman-Freedman A., Jacobs WR J.r., Chan J., Porcelli S.A., Achkar J.M, & Casadevall A. (2017). Enhanced control of Mycobacterium tuberculosis extrapulmonary dissemination in mice by an arabinomannan-protein conjugate vaccine. PLoS Pathogens, 13(3), e1006250.

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Immunohistochemical Analysis of Intestinal Tissue

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Immunostaining was performed similar to previously described (Huffman, et al. 2007 (link); Huffman, et al. 2008 (link)). In brief, intestinal sections were subjected to antigen retrieval (Citrate buffer pH 6) using a pressure cooker on high steam for 10 min. Following rehydration, slides were treated with 3.0% H₂O₂ for 5 min to quench endogenous peroxidase activity, subjected to an avidin-biotin blocking step (Vector Labs SP-2001), and subsequently blocked with preimmune goat or rabbit serum (1%) for 20 min. Sections were then incubated with an antibody against Ki67 (1:400; cat#12202) pAkt^Ser473 (1:50; cat#4060), phospho- (Ser/Thr) Akt Substrate (1:500; cat#9611), β-catenin (1:100; cat#8480), and anti-BrdU antibody (1:200; cat#5292) from Cell Signaling. A negative control was included in the same run using a subset of slides by omitting primary antibody from the staining procedure. Sections were then incubated with the appropriate biotinylated secondary antibody for 20 min, followed by a streptavidin-HRP detection system (Vector) and application of 3,3′-diaminobenzidine (DAB) for visualization of the antigen-antibody complex (Scytek). Digital files of all slides were then acquired with a PerkinElmer P250 High Capacity Slide Scanner and positive stained cells were analyzed using QuantCenter Software.

Tabrizian T., Wang D., Guan F., Hu Z., Beck A.P., Delahaye F, & Huffman D.M. (2017). Apc inactivation, but not obesity, synergizes with Pten deficiency to drive intestinal stem cell-derived tumorigenesis. Endocrine-related cancer, 24(6), 253-265.

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Lung Metastasis Quantification Protocol

Cited 1 time

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All experiments were performed according to protocols approved by the Animal Welfare Committee at the Albert Einstein College of Medicine. MDA-MB-231 cells (5 × 10⁵ cells/100 μl per mouse) were injected into the lateral tail vein of 8-week old female SCID mice. After six or eight weeks, the mice were euthanized. The lungs were excised, fixed in 10% buffered formalin, paraffin embedded, sectioned, and stained with hematoxylin and eosin or immunostained with a human- specific pan-cytokeratin antibody (4.5 μg/ml, Leica cat #AE1/AE-L-CE) using DAB detection. The slides were scanned using a Perkin Elmer P250 High Capacity Slide Scanner. The metastatic lesions in lungs 6 weeks after cell injection were analyzed by tracing the lungs and me- tastases using 3D Histech CaseViewer 1.4 software. The metastatic lesions in lungs 8 weeks after cell injection were analyzed using image analysis software developed for DAB immunohistochemistry [29 (link)].

Norwood Toro L.E., Wang Y., Condeelis J.S., Jones J.G., Backer J.M, & Bresnick A.R. (2018). Myosin-IIA heavy chain phosphorylation on SI943 regulates tumor metastasis. Experimental cell research, 370(2), 273-282.

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Measuring Apoptosis-Induced DNA Damage

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Apoptosis induced DNA damage was measured using the ApopTag® Peroxidase In Situ Apoptosis Detection Kit (S7100, EMD Millipore, Temecula, CA) following the manufacturer’s protocol for paraffin-embedded tissue. Testis sections were deparaffinized using Histo-Clear reagent (HS-200, National Diagnostics, Atlanta, GA). Stained slides were scanned using a Perkin Elmer P250 high capacity slide scanner and images were analyzed using FIJI software to count foci^{38 (link)}.

Akintayo A., Liang M., Bartholdy B., Batista F., Aguilan J., Prendergast J., Sabrin A., Sundaram S, & Stanley P. (2020). The Golgi Glycoprotein MGAT4D is an Intrinsic Protector of Testicular Germ Cells From Mild Heat Stress. Scientific Reports, 10, 2135.

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Tissue Preparation and Imaging Protocol

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Tissue samples were fixed in 10% neutral buffered formalin and processed for hematoxylin and eosin staining. For each organ collected and for each genotype, two sections were cuts at 100 μm apart and all slides were scanned using a P250 High Capacity Slide Scanner (Perkin Elmer).

Chorro L., Suzuki M., Chin S.S., Williams T.M., Snapp E.L., Odagiu L., Labrecque N, & Lauvau G. (2018). Interleukin 2 modulates thymic-derived regulatory T cell epigenetic landscape. Nature Communications, 9, 5368.

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Quantifying Lung Pathology in Mice

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Lungs were removed and fixed in 10% neutral buffered formalin (Fisher Scientific, Fair Lawn, NJ). Tissues were embedded with paraffin, sectioned at 5-μm thickness, and stained with hematoxylin and eosin. Five different lung sections per mouse were analyzed. Slides were scanned with a Perkin-Elmer P250 high-capacity slide scanner (Waltham, MA) at 2,000 dots per inch (dpi). Digitized images were then analyzed using ImageJ software to calculate the total disease area occupied by granuloma and the percentage of lung surface affected by pneumonia as well as the number of infiltrates per lung. The total disease area for the entire lung section was measured by adding the values for each lesion. The total percentage of diseased tissue was calculated by dividing the total disease area by the entire lung section and multiplying by 100, using ImageJ software.

Kumagai T., Palacios A., Casadevall A., García M.J., Toro C., Tiemeyer M, & Prados-Rosales R. (2019). Serum IgM Glycosylation Associated with Tuberculosis Infection in Mice. mSphere, 4(2), e00684-18.

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Quantitative Analysis of Apoptosis in MOLM-13 Xenografts

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Femur from MOLM-13 xenografts of mice treated with 10 mg/kg BTSA1 or vehicle for 3 weeks were dissected and fixed in 10% buffered formalin (Fisher Scientific) for 24 hr followed by decalcification of bone. Paraffin-embedded sections were then stained with cleaved caspase-3 antibody (Cell Signaling Cat. 9661) or with the ApopTag Peroxidase in situ Apoptosis Detection Kit (Millipore Cat. S7100), which detects apoptotic cells in situ by labeling and detecting DNA strand breaks by the TUNEL method. Stained slides were imaged using the PerkinElmer P250 High Capacity Slide Scanner under brightfield using a 20X objective. Quantification and analysis of densitometry of immunohistochemistry (IHC) was performed using Panoramic Viewer. Using Panoramic Viewer, bone marrow sections from slides were selected followed by analysis using the “Densito Quant Module” This module, measures the density of immunostain on the digital slides by distributing pixels to negative and 3 grades of positive classes by their RGB values. For caspase cleavage stain, the percentage of total positive signals of BTSA1 or vehicle treated samples was obtained using the Densito Quant Module and plotted as bar-graphs. For TUNEL stain, the percentage the of total medium and strong positive signals of BTSA1 or Vehicle treated samples was obtained using the Densito Quant Module and plotted as bar-graphs.

Reyna D.E., Garner T.P., Lopez A., Kopp F., Choudhary G.S., Sridharan A., Narayanagari S.R., Mitchell K., Dong B., Bartholdy B.A., Walensky L.D., Verma A., Steidl U, & Gavathiotis E. (2017). Direct activation of BAX by BTSA1 overcomes apoptosis resistance in acute myeloid leukemia. Cancer cell, 32(4), 490-505.e10.

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Gastrointestinal Tract Histological Analysis

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Evaluation of the gastrointestinal tract was performed following isolation and division into four segments: duodenum, jejunum, ileum and colon, as previously described (6 (link), 36 (link)). Tissues were rolled and fixed overnight in 10% neutral-buffered formalin at 4°C, processed through a series of alcohols and xylenes, and embedded in paraffin. Hematoxylin & Eosin (H&E) stained sections (5 μm) from each segment of small intestine were then scanned into digital files with a PerkinElmer P250 High Capacity Slide Scanner, and crypt area and villi length were evaluated in Panoramic Viewer using 50 random villi per animal to form a single composite average.

Guan F., Tabrizian T., Novaj A., Nakanishi M., Rosenberg D.W, & Huffman D.M. (2018). Dietary Walnuts Protect Against Obesity-Driven Intestinal Stem Cell Decline and Tumorigenesis. Frontiers in Nutrition, 5, 37.

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